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Related Experiment Videos

Rhodopsin in the rod outer segment plasma membrane.

S Basinger, D Bok, M Hall

    The Journal of Cell Biology
    |April 1, 1976
    PubMed
    Summary

    Newly synthesized visual pigment is found in frog rod outer segments, including basal disks and plasma membranes. Protein synthesis inhibition blocks this visual pigment incorporation into these structures.

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    Area of Science:

    • Biochemistry
    • Cell Biology
    • Neuroscience

    Background:

    • The rod outer segment (ROS) is the photoreceptor cell's light-sensing organelle.
    • Understanding the synthesis and localization of visual pigment is crucial for photoreceptor function.

    Purpose of the Study:

    • To investigate the synthesis, localization, and configuration of visual pigment in isolated frog retinas.
    • To identify the newly synthesized protein in rod outer segments.

    Main Methods:

    • In vitro incubation of isolated frog retinas with [3H]leucine pulse-chase.
    • Light and electron microscope autoradiography.
    • Biochemical analysis of purified outer segments using SDS-PAGE and gel filtration chromatography.

    Main Results:

    • Autoradiograms showed intense radioactivity in basal disks and plasma membranes of rod outer segments.
    • Puromycin significantly inhibited labeling, indicating protein synthesis dependence.
    • Biochemical analysis revealed that approximately 90% of the radioactivity co-migrated with visual pigment.
    • Newly synthesized visual pigment was found in both basal disks and plasma membrane in its native configuration.

    Conclusions:

    • The primary newly synthesized protein in frog rod outer segments is visual pigment.
    • This visual pigment is incorporated into both basal disks and the plasma membrane.
    • The newly synthesized visual pigment maintains its native configuration within these structures.

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